Assays for Follicle Stimulating Hormone and Luteinising Hormone: Guidelines for the Provision of a Clinical Biochemistry Service

Abstract

The measurement of serum follicle stimulating hormone (FSH) and luteinising hormone (LH), together with the appropriate sex steroid, is of great value in the investigation of delayed and precocious puberty, hypogonadism, subfertility, polycystic ovarian disease and hypothalamic-pituitary disorders. Dynamic function testing of the hypothalamic-pituitary-gonadal axis should be restricted to a few defined situations. Sequential LH measurements, either in serum or in urine, may be used to time ovulation during artificial insemination or in vitro fertilisation programmes. No special precautions are necessary when sampling for FSH and LH measurement; serum is preferred to plasma and should be stored frozen before assay. Aliquots of timed urine specimens of known volume should be stored frozen without preservative. Gonadotropin results should be available within 2–3 weeks; laboratories unable to meet this schedule are advised to send their samples to a Regional Centre for assay. Reagents for the radioimmunoassay of FSH and LH are readily available, and standard techniques have been developed for their use. Laboratories using ‘in-house’ methods should pay particular attention to the matrix used for preparing standard solutions, the purification of radioligands and the optimisation of the separation system. Low cost matched reagents of proven performance are available in kit form from the Chelsea Hospital for Women; several commercial kits are also available, although few are widely used in the UK. The overall performance of laboratories in the UK External Quality Assessment Scheme (EQAS) for FSH and LH has remained steady for several years. Of the 130 participants, only about 15% in each scheme have ‘good’ performance (cumulative bias less than 10%, plus cumulative variability of bias less than 10%), whilst a similar proportion have ‘unacceptable’ performance (cumulative bias greater than 20% and/or cumulative variability of bias greater than 25%). The remaining 70% of laboratories have ‘adequate’ performance but are at risk of producing results that are clinically misleading. Within any one method group, the performance of FSH and LH assays are closely related. Optimal assay performance depends upon sensible laboratory management to ensure skilled operators, a regular programme of reagent/kit renewal, comprehensive internal and external quality assessment, and attention to detail in all aspects of gonadotrophin assay. The working range of each individual assay should be defined and no absolute result reported from outside this range. Mean intra-assay and interassay coefficients of variation on selected human serum quality control pools should be better than 8% and 15%, respectively, for both gonadotrophins. All laboratories performing FSH and LH assays should belong to the UK EQAS for gonadotrophins. Immunometric assays, using monoclonal antibodies, will supersede radioimmunoassays for FSH and LH during the next few years. Some of these assays will have non-isotopic labels.